HemaTox™ Megakaryocyte Kit includes a specialized serum-free culture medium and 100X supplement. Complete HemaTox™ Megakaryocyte Medium (HemaTox™ Megakaryocyte Medium + HemaTox™ Megakaryocyte 100X Supplement) promotes the proliferation of human CD34+ HSPCs and their differentiation into megakaryocytes during a 10-day culture period. After culture, the cells can be counted and assessed for expression of megakaryocyte marker CD41 using flow cytometry or other methods.
HemaTox™ Megakaryocyte Kit may be used on its own or in combination with HemaTox™ Myeloid Kit (Catalog #09704) or HemaTox™ Erythroid Kit (Catalog #09701) to assess lineage-specific drug toxicity in parallel.
Each kit contains sufficient medium and supplement for testing up to 160 different conditions (triplicate wells per condition, 200 µL per well) in 5 x 96-well plates.
- HemaTox™ Megakaryocyte Medium, 100 mL
- HemaTox™ Megakaryocyte 100X Supplement, 1 mL
Data and Publications
Figure 1. General HemaTox™ Megakaryocyte Kit Procedure
*The cell isolation step may be omitted if pre-enriched CD34+ cells are used.
Figure 2. Flow Cytometry Plot Showing Cells Produced After Culture of CD34+ HSPCs with the HemaTox™ Megakaryocyte Kit
(A) Human CB CD34+ cells were cultured with the HemaTox™ Megakaryocyte Kit using the protocol as written in the Product Information Sheet (PIS). (B) After the appropriate culture period, cells were harvested and stained for cell surface proteins expressed on megakaryocyte (CD41 and CD45) cells.
Figure 3. Reproducibility of HemaTox™ Megakaryocyte Kit Results Between Experiments and Using Different CD34+ Cell Preparations
(A) Dose-response curves were generated from titrations of 5-FU added to human CB CD34+ cells isolated from 3 donors and cultured with the HemaTox™ Megakaryocyte Kit. Three to five separate experiments were performed with cells from each donor. In each assay similar IC50 values were obtained with cells from different donors and in different experiments with cells from the same donor. Shown are values normalized to the percentages (%) of maximum cell growth without drug. Despite differences in absolute cell counts, curves are reproducible when normalized within each experiment. (B) Table showing IC50 values generated for 5-FU in culture with the megakaryocyte kit including standard deviation (SD) and the coefficient of variation (% CV) across experiments.
Figure 4. Lineage-Specific Differences in Hematotoxicity Identified with HemaTox™ Erythroid, Myeloid and Megakaryocyte Kits
Results show average IC50 values for each drug tested on human BM CD34+ cells using the HemaTox™ Erythroid (grey), Myeloid (gold) and Megakaryocyte (orange) Kits. Most drugs show similar toxicity for each lineage but some, such as Sunitinib, are ~100-fold more toxic for erythroid than for megakaryocyte progenitor differentiation with intermediate toxicity for myeloid progenitor differentiation. Vertical lines indicate standard error of the mean (SEM) (n = 4 - 8).